High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity

Welter A, Sundararaman S, Li R, Zhang T, Karulin AY, Lehmann A, Naeem V, Roen DR, Kürten S, Lehmann PV (2018)

Publication Type: Journal article

Publication year: 2018

Journal

Cells MDPI

Book Volume: 7

Journal Issue: 5

DOI: 10.3390/cells7050035

Abstract

One of the primary effector functions of immune cells is the killing of virus-infected or malignant cells in the body. Natural killer (NK) and CD8 effector T cells are specialized for this function. The gold standard for measuring such cell-mediated cytolysis has been the chromium release assay, in which the leakage of the radioactive isotope from damaged target cells is being detected. Flow cytometry-based single cell analysis of target cells has recently been established as a non-radioactive alternative. Here we introduce a target cell visualization assay (TVA) that applies similar target cell staining approaches as used in flow cytometry but based on single cell computer image analysis. Two versions of TVA are described here. In one, the decrease in numbers of calcein-stained, i.e., viable, target cells is assessed. In the other, the CFSE/PI TVA, the increase in numbers of dead target cells is established in addition. TVA assays are shown to operate with the same sensitivity as standard chromium release assays, and, leaving data audit trails in form of scanned (raw), analyzed, and quality-controlled images, thus meeting requirements for measuring cell-mediated cytolysis in a regulated environment.

Authors with CRIS profile

Stefanie Kürten Lehrstuhl für Mikroskopische Anatomie und Molekulare Zellbiologie

Involved external institutions

CTL Europe GmbH DE Germany (DE)

How to cite

APA:

Welter, A., Sundararaman, S., Li, R., Zhang, T., Karulin, A.Y., Lehmann, A.,... Lehmann, P.V. (2018). High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity. Cells, 7(5). https://dx.doi.org/10.3390/cells7050035

MLA:

Welter, Anna, et al. "High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity." Cells 7.5 (2018).

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